Knockdown of PKD1 Decreased Sensitivity of Human Salivary Gland Adenoid Cystic Carcinoma Cell Line ACC2 to Paclitaxel
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摘要:目的
研究蛋白激酶D（PKD）沉默对人涎腺腺样囊性癌细胞ACC-2细胞增殖、迁移、化疗药物敏感度及凋亡的影响。
方法转染Control-shRNA和PKD1-shRNA质粒后，药物筛选出稳定转染的ACC-2细胞系，并用Western blot验证细胞中PKD1敲除效率；划痕实验检测PKD1敲除后细胞迁移能力改变；CCK-8法检测PKD1敲除后细胞增殖能力以及紫杉醇对细胞的半致死浓度变化；PI染色并用流式细胞仪检测紫杉醇处理并PKD1敲除后细胞凋亡情况变化。
结果建立了PKD1基因沉默的稳定细胞株；相较于对照组，实验组PKD1-sh细胞的增殖能力和迁移能力没有显著变化，但紫杉醇半致死浓度增高，紫杉醇处理后的细胞凋亡率降低。
结论PKD1沉默降低了ACC-2细胞对紫杉醇的药物敏感度，抑制了紫杉醇引起的细胞凋亡。
Abstract:ObjectiveTo explore the effect of silencing PKD1 on the proliferation, migration, chemosensitivity and apoptosis of human salivary gland adenoid cystic carcinoma cell line ACC-2.
MethodsACC2 cell lines were transfected with either the Control-shRNA or PKD1-shRNA plasmids. The stable transfected cells were selected using puromycin and the efficiency of PKD1 knockdown was detected by Western blot. The cell migration of each cell lines was detected using wound-healing assay. The growth ability and the 50% inhibitory concentrations (IC50) of paclitaxel of the control and PKD1 knockdown cell lines were detected using Cell Counting Kit-8(CCK-8) . After by paclitaxel treatment, the control and PKD1 knockdown cell lines were stained using propidium iodide(PI) and the apoptosis cell rates were measured by flow cytometry.
ResultsThe stable ACC-2 cell line with PKD1 knockdown were established. Compared with the parental cells, the proliferation and migration of PKD1-sh cells were not significantly different, but their resistance to paclitaxel had a significant increase, evident as the increase in IC50 value and the decrease in cell apoptosis rate.
ConclusionThe shRNA silencing of PKD1 in ACC-2 cells decreases their chemosensitivity to paclitaxel and inhibits paclitaxel-induced cell apoptosis.
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Key words:
- PKD /
- Paclitaxel /
- Drug resistance
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图 1 构建稳定敲除PKD1的ACC-2细胞系
Figure 1 Establishment of a stable PKD1-knockdown ACC-2 cell line
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图 4 PKD1敲除使细胞对紫杉醇抗性增加
Figure 4 Resistance to paclitaxel was increased by PKD1 knockdown
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